how to label all proteins after western-blot - someting like ponceau red, but after blocking (Jul/13/2006 )

Hello there,

I did a western-blot after biotinylated proteins capture. I still cannot see the band I'm interested in. I"m wondering if I managed to capture the proteins or not.

I have a question : is there a way to color all proteins on a nitrocellulose membrane, after western-blotting? (i.e. after blocking and incubating antibodies)

I guess that I will have troubles because of blocking with 3% BSA and milk 1%.
I tried once with coomassie blue, because a collegue of mine manage to color his PVDF membrane after western-blot, but my nitrocellulose membrane become simply completely blue.

May be I should strip and try ponceau red, or coomassie?

Thanks in advance

Missele

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hi
if you want stgh to stain proteins, you'll surely label milk proteins and BSA. so the whole membrane will be red. So i think that youl'' better have to strip the membrane and stain with ponceau red

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i think this is the solution if u do stain ur whole membrane it will stain all the proteins in ur case( blocking\).

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Thank you spanishflower and Fred,

i will strip, no miracle.
lucky guys the one who use PVDF, it works even if you block !
but nitrocellulose has other advantages

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woaw "la nuit porte conseil", it means night gives you good advie.
however I realize I only wake up at 10:00 am
it's biotinylated proteins. I will put some streptavidin-HRP and detect them !
Sorry again for asking you such a stupid question.

(I really need holidays)

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mais le matin est plus malin...
since you are able to adjust your exp, there is no problem. And there is no stupid question....

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I didn't know this one.

Thanks Fred.

Ok then , I think it's time to go home. I'll be more smart tomorrow morning !
bye

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